Antoni J. Duleba

Expectant management of choriocarcinoma limited to placenta

Choriocarcinoma limited to placenta was discovered incidentally following a seemingly uneventful term pregnancy. The newborn had unsuspected severe anemia and thrombocytopenia, due to fetomaternal hemorrhage. His recovery was good following transfusions. The mother was asymptomatic and her extensive workup for metastatic choriocarcinoma was negative. She was subsequently managed expectantly and monitored with serial serum beta-hCG, demonstrating near-logarithmic decline to non-pregnant levels within 5 weeks of delivery.

Direct effects of clomiphene citrate on the steroidogenic capability of human granulosa cells

The effects of clomiphene citrate (CC), 17 beta-estradiol (E2), and human chorionic gonadotropin (hCG) on the accumulation of progesterone (P) and 20 alpha-hydroxy-4-pregnen-3-one (20 alpha-OHP) in cultured human granulosa cells (GC) were examined. In addition, the metabolism of [4-14C]pregnenolone and accumulation of [4-14C]P in response to CC and E2 were determined. The authors conclude the following: (1) the dose-dependent inhibition of P and 20 alpha-OHP production by CC in GC was not reproduced by E2, (2) hCG abolished these effects of CC, (3) these inhibitory actions of CC were not associated with altered 3 beta-hydroxysteroid dehydrogenase activity nor P catabolism indicating that, (4) these actions by CC on the GC occur at some step(s) during steroidogenesis preceding the formation of pregnenolone. These findings may explain, at least in part, the luteal deficiency experienced by women treated with CC, and they also provide a rationale for the use of hCG supplementation during ovulation induction with CC.

Effect of cannabinoids on progesterone production by ovarian granulosa cells of pig and rat

Abstract Various cannabinoids inhibited both basal and FSH-stimulated progesterone synthesis by pig and rat granulosa cells in vitro in a dose- (0.09 – 16 μM) and time- (4 – 24 h) dependent manner. The data present further evidence for a possible direct interference of cannabinoids with ovarian functions.

Effects of prostaglandins E2 and F2α on progesterone metabolism by rat granulosa cells

Abstract Rat granulosa cells were cultured with or without PGE 2 and/or PGF 2α . Accumulation of endogenous progesterone and 20α-hydroxy-4-pregnen-3-one was determined. Additionally, [4- 14 C]progesterone metabolism was assessed. PGE 2 increased progesterone accumulation, in part, by decreasing progesterone catabolism to 20α-reduced progestins. In contrast, PGF 2α stimulated 20α-hydroxysteroid dehydrogenase activity, thus increasing progesterone catabolism. Combined treatment with PGE 2 and PGF 2α augmented progesterone accumulation to levels above controls but below those attained with PGE 2 alone. These data indicate that PGE 2 and PGF 2α exert opposite effects on progesterone production and catabolism and that the ratio of PGE 2 to PGF 2α in the local granulosa cell milieu may be of importance in determining overall progesterone output.

Differential actions of FSH and androgens on progesterone catabolism by rat granulosa cells1

Rat granulosa cells were preincubated with follicle-stimulating hormone (FSH, 1 micrograms/ml) and/or androgens (testosterone and 5 alpha-dihydrotestosterone, 0.5 microM) and subsequent [4-14C]progesterone metabolism by these cells was studied. Granulosa cells metabolized radiolabeled progesterone to three major products 20 alpha-hydroxy-4-pregnen-3-one, 3 alpha-hydroxy-5 alpha-pregnan-20-one, and 5 alpha-pregnane-3 alpha, 20 alpha-diol. Androgens, but not FSH, decreased overall progesterone utilization. Both FSH and androgens decreased conversion of progesterone to 20 alpha-hydroxy-4-pregnen-3-one. The accumulation of 5 alpha-pregnane-3 alpha, 20 alpha-diol was increased by FSH, decreased by testosterone, and not altered by 5 alpha-dihydrotestosterone. The accumulation of 3 alpha-hydroxy-5 alpha-pregnan-20-one was significantly increased by both FSH and androgens. Present results indicate that FSH as well as androgens modulate progesterone metabolism by rat granulosa cells. FSH appears to increase 5 alpha-reductase activity and/or decrease 20 alpha-hydroxysteroid dehydrogenase activity while androgens decrease only 20 alpha-hydroxysteroid dehydrogenase activity.

Androgenic alteration in pathways of C19-steroid metabolism by cultured rat granulosa cells

Pretreatment of rat granulosa cells with androgens altered the subsequent metabolism of [4-14C]testosterone in vitro. In cells pretreated with testosterone or 5 alpha-dihydrotestosterone, the accumulation of radiolabeled 5 alpha-androstane-3 alpha, 17 beta-diol decreased by 40% and 36%, respectively, when compared with control, while the levels of androstenedione increased by 64% and 54%, respectively. For comparison, pretreatment with 0.5 microM 17 beta-estradiol failed to affect the level of radioactivity incorporated into all three major metabolites. In other experiments, granulosa cells were incubated in the presence of 0.5 microM [4-14C]dihydrotestosterone. While 5 alpha-androstane-3 alpha, 17 beta-diol was decreased slightly by pretreatment with testosterone or 5 alpha-dihydrotestosterone, radiolabeled androsterone was increased significantly. These data suggest a possible modulatory action of androgens on ovarian 17 beta-hydroxysteroid dehydrogenase and/or 5 alpha-reductase activity.

Comparative studies of androgen metabolism in theca and granulosa cells of human follicles in vitro

In eight separate experiments, theca and granulosa were isolated from human follicles (5-25 mm in diameter), and their capacities to metabolize radiolabelled testosterone in 24 hour cultures were assessed. Theca metabolized testosterone primarily to androstenedione, however significant aromatization to estradiol-17 beta and to estrone was also observed. Granulosa metabolized testosterone primarily to estra-diol-17 beta and estrone, while smaller quantities were converted to androstenedione. In seven of these experiments, the intermediate of aromatization, 19-hydroxytestosterone, was identified. In six of these experiments, theca, when compared to granulosa, produced more androstenedione but less estradiol-17 beta and estrone. 5 alpha-Reduced androgens were non-detectable or produced in small quantities. In a single experiment, metabolism of androstenedione was compared to metabolism of testosterone by both theca and granulosa. Theca metabolized androstenedione to testosterone in smaller quantities than testosterone to androstenedione. Granulosa metabolized androstenedione to testosterone in higher quantities than testosterone to androstenedione. Both theca and granulosa aromatized androstenedione more readily than testosterone.

Androgenic modulation of progesterone metabolism by rat granulosa cells in culture

Effects of androgens on progesterone accumulation, utilization of exogenous progesterone and accumulation of [4-14C]progesterone metabolites by rat granulosa cells in culture were studied. Androgen increased progesterone accumulation in cultures without exogenous progesterone and slowed the overall decline of progesterone concentration in cultures supplemented with exogenous progesterone. Both aromatizable testosterone and nonaromatizable 5 alpha-dihydrotestosterone decreased [4-14C]progesterone utilization by granulosa cells by 12 to 30%. This effect was observed irrespective of whether the cells were continuously exposed to androgens or only pre-exposed. In he same experiments, androgens decreased conversion of radiolabeled progesterone to 20 alpha-hydroxy-4-pregnen-3-one by 11 to 50% and to 5 alpha-pregnane-3 alpha, 20 alpha-diol by 26 to 49%. Accumulation of 3 alpha-hydroxy-5 alpha-pregnan-20-one was not altered in 3 h incubations and was increased by up to 43% in 24 h incubations by androgen treatment. It is suggested that androgens alter progesterone catabolism by granulosa cells by decreasing 20 alpha-hydroxysteroid dehydrogenase activity and that this effect may contribute to overall stimulatory action of androgens on progesterone accumulation.

Time-dependent effects of follicle-stimulating hormone on progesterone metabolism by cultured rat granulosa cells

The effects of FSH on the accumulation of endogenous progesterone and 20 alpha-hydroxy-4-pregnen-3-one as well as on the metabolism of [4-14C]progesterone were studied in 24, 48 and 72 h cultures of rat granulosa cells. FSH stimulated the accumulation of both progesterone and 20 alpha-hydroxy-4-pregnen-3-one by 6-18 fold and 2.5-44 fold, respectively. Short term exposure (24 h) to FSH resulted in the ratio of progesterone to 20 alpha-hydroxy-4-pregnen-3-one accumulations significantly increased (by 2.6 fold), while the reverse was observed for the longer (48 and 72 h) cultures whereby control levels of the ratio of progesterone to 20-alpha-hydroxy-4-pregnen-3-one were significantly greater (by 50% and 270%, respectively) than those of FSH-treated cultures. Effects of FSH on [4-14C]progesterone were also time-dependent. Twenty-four hour cultures were associated with FSH induced inhibition of 20 alpha-reduced metabolites of progesterone (by 55%) while the 48 and 72 h exposures to FSH resulted in a significant increase of the 20 alpha-reduced metabolites above control levels (by 73% and 230%, respectively). Consequently, it is postulated that FSH may exert biphasic time-dependent actions on 20 alpha-hydroxysteroid dehydrogenase activity with short term inhibitory and longer term stimulatory effects.

Actions of LH/hCG on accumulation and catabolism of progestins in cultured rat granulosa cells

Immature hypophysectomized rats were treated with estradiol-17 beta and follicle-stimulating hormone. Granulosa cells were isolated and incubated for 24 h with or without varying doses of ovine luteinizing hormone (NIMADD-oLH-24) or human chorionic gonadotropin (NIADDK CR 125) and accumulations of progesterone and 20 alpha-hydroxy-4-pregnen-3-one were determined. The cells were reincubated for 3 h with [4-14C]progesterone (0.5 nmol/mL) and the radiolabelled metabolites were separated and quantified. Both LH (0.04-1.0 ug/mL) and hCG (0.04-1.0 ug/mL) enhanced the accumulation of endogenous progesterone (by up to 300 and 150%, respectively) and 20 alpha-hydroxy-4-pregnen-3-one (by up to 90 and 85%, respectively) producing dose-dependent increases of the ratio of progesterone to 20 alpha-hydroxy-4-pregnen-3-one (by up to 125 and 70%, respectively). Studies of the metabolism of [1-14C] progesterone have demonstrated that both LH and hCG led to a dose-dependent decrease of the utilization of radiolabelled progesterone (down to 64 and 70%, respectively, of the control value). This effect was associated with an LH- and hCG-dependent inhibition of 20 alpha-hydroxysteroid dehydrogenase activity (down to 60 and 70%, respectively, of the control value) but had no significant effect on 5 alpha-reductase. The present results indicate that LH and hCG stimulate accumulation of progesterone at least in part by decreasing the 20 alpha-hydroxysteroid dehydrogenase activity.